Methods for generating therapeutic delivery platforms

Abstract

Methods for producing engineered exosomes and other vesicle-like biological targets, including allowing a target vesicle-like structure to react and bind with immunomagnetic particles; capturing the immunomagnetic particle/vesicle complex by applying a magnetic field; further engineering the captured vesicles by surface modifying with additional active moieties or internally loading with active agents; and releasing the engineered vesicle-like structures, such as by photolytically cleaving a linkage between the particle and engineered vesicle-like structures, thereby releasing intact vesicle-like structures which can act as delivery vehicles for therapeutic treatments.

The invention claimed is: 
     
       1. A composition for isolating an extracellular vesicle, the composition comprising a plurality of magnetic particles, each magnetic particle comprising:
 a core particle, wherein said core particle is coated with a layer comprising graphene-oxide nanosheets covalently bonded to the core particle, further comprising at least one polydopamine polymer coupled with the graphene-oxide nanosheets; and 
 a targeting moiety extending from the core particle via a photocleavable linker, wherein said targeting moiety is coupled to at least one polydopamine polymer, and said photocleavable linker connecting the magnetic particle and the targeting moiety, wherein the targeting moiety specifically binds to an extracellular vesicle surface marker for isolating said extracellular vesicle. 
 
     
     
       2. The composition of  claim 1 , wherein said magnetic particles are dispersed in a buffer solution. 
     
     
       3. The composition of  claim 1 , wherein said targeting moiety is an antigenic peptide, antibody or a portion thereof, aptamer, antigenic epitope, affinity peptide, cell surface receptor ligand, or MHC binding peptide. 
     
     
       4. The composition of  claim 1 , wherein the targeting moiety specifically binds to an extracellular vesicle surface marker selected from the group consisting of CD9, CD63, and CD81. 
     
     
       5. The composition of  claim 1 , wherein the targeting moiety specifically binds to CD9. 
     
     
       6. The composition of  claim 1 , wherein the targeting moiety specifically binds to CD63. 
     
     
       7. The composition of  claim 1 , wherein the targeting moiety specifically binds to CD81. 
     
     
       8. The composition of  claim 1 , wherein the targeting moiety specifically binds to an extracellular vesicle surface marker selected from the group consisting of an MHC class I molecule, an MHC class II molecule, an interleukin, TNFα, IFNγ, RANTES, G-CSF, M-CSF, IFNα, CTAPIII, ENA-78, GRO, I-309, PF-4, IP-10, LD-78, MGSA, MIP-1α, MIP-1β, and combinations thereof. 
     
     
       9. The composition of  claim 1 , wherein said magnetic particle has a diameter of at least 500 nm. 
     
     
       10. The composition of  claim 1 , wherein said magnetic particle has a diameter of about 500 nm to about 1000 nm. 
     
     
       11. The composition of  claim 1 , each magnetic particle comprising said core particle and a plurality of targeting moieties extending from the core particle via respective photocleavable linkers. 
     
     
       12. The composition of  claim 1 , wherein the photocleavable linker comprises a biotinylated end bound to an avidin, streptavidin, or neutravidin coating on said core particle, a cleavable linking group that is cleaved upon exposure to a wavelength of light, and a functional end bound to said targeting moiety. 
     
     
       13. The composition of  claim 12 , wherein said wavelength ranges from 100 to 380 nm. 
     
     
       14. The composition of  claim 12 , wherein said photocleavable linker is a linear chain. 
     
     
       15. The composition of  claim 12 , wherein said functional end comprises an amine moiety for attachment to targeting moiety. 
     
     
       16. The composition of  claim 12 , cleavable linking group is selected from the group consisting of 6-nitroveratryloxycarbonyl (NVOC), 2-nitrobenzyloxycarbonyl (NBOC), α,α-dimethyl-dimethoxybenzyloxycarbonyl (DDZ), ortho-nitrobenzyl (ONB), 1-(2-nitrophenyl)ethyl (NPE), alpha-carboxy-2-nitrobenzyl (CNB), 4,5-dimethoxy-2-nitrobenzyl (DMNB), 1-(4,5-dimethoxy-2-nitrophenyl)ethyl (DMNPE), 5-carboxymethoxy-2-nitrobenzyl (CMNB), (5-carboxymethoxy-2-nitrobenzyl)oxy)carbonyl (CMNCBZ), nitrodibenzofurane, and substituted derivatives thereof. 
     
     
       17. The composition of  claim 16 , wherein said substituted derivatives comprise a substitution on an aromatic ring. 
     
     
       18. The composition of  claim 12 , wherein said photocleavable linker comprises 
       
         
           
           
               
               
           
         
         wherein the dashed line indicates the bond cleaved during exposure to light. 
       
     
     
       19. The composition of  claim 1 , wherein said extracellular vesicle is an exosome.